Laboratorio de Inmunología y Virología, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León, Ciudad Universitaria, C,P, 66450, San Nicolás de los Garza, Nuevo León, México.
Virol J. 2012 Dec 12;9:307. doi: 10.1186/1743-422X-9-307.
Newcastle Disease Virus (NDV) causes a serious infectious disease in birds that results in severe losses in the worldwide poultry industry. Despite vaccination, NDV outbreaks have increased the necessity of alternative prevention and control measures. Several recent studies focused on antiviral compounds obtained from natural resources. Many extracts from marine organisms have been isolated and tested for pharmacological purposes, and their antiviral activity has been demonstrated in vitro and in vivo. Fucoidan is a sulfated polysaccharide present in the cell wall matrix of brown algae that has been demonstrated to inhibit certain enveloped viruses with low toxicity. This study evaluated the potential antiviral activity and the mechanism of action of fucoidan from Cladosiphon okamuranus against NDV in the Vero cell line.
The cytotoxicity of fucoidan was determined by the MTT assay. To study its antiviral activity, fusion and plaque-forming unit (PFU) inhibition assays were conducted. The mechanism of action was determined by time of addition, fusion inhibition, and penetration assays. The NDV vaccine strain (La Sota) was used in the fusion inhibition assays. PFU and Western blot experiments were performed using a wild-type lentogenic NDV strain.
Fucoidan exhibited antiviral activity against NDV La Sota, with an obtained IS50 >2000. In time of addition studies, we observed viral inhibition in the early stages of infection (0-60 min post-infection). The inhibition of viral penetration experiments with a wild-type NDV strain supported this result, as these experiments demonstrated a 48% decrease in viral infection as well as reduced HN protein expression. Ribavirin, which was used as an antiviral control, exhibited lower antiviral activity than fucoidan and high toxicity at active doses. In the fusion assays, the number of syncytia was significantly reduced (70% inhibition) when fucoidan was added before cleavage of the fusion protein, perhaps indicating a specific interaction between fucoidan and the F0 protein.
The results of this study suggest that fucoidan from C. okamuranus represents a potential low-toxicity antiviral compound for the poultry industry, and our findings provide a better understanding of the mode of action of sulfated polysaccharides.
新城疫病毒（NDV）会导致禽类发生严重的传染病，从而使全球家禽业遭受严重损失。尽管进行了疫苗接种，但 NDV 的爆发增加了对替代预防和控制措施的需求。最近的几项研究侧重于从天然资源中获得的抗病毒化合物。已经从海洋生物中分离出许多提取物并用于药理学目的，并且已经在体外和体内证明了它们的抗病毒活性。褐藻中的硫酸多糖岩藻聚糖已被证明可抑制某些包膜病毒，且毒性低。本研究评估了来自 Cladosiphon okamuranus 的褐藻聚糖对 Vero 细胞系中 NDV 的潜在抗病毒活性和作用机制。
通过 MTT 测定法测定褐藻聚糖的细胞毒性。为了研究其抗病毒活性，进行了融合和蚀斑形成单位（PFU）抑制测定。通过添加时间、融合抑制和渗透测定来确定作用机制。融合抑制测定中使用了 NDV 疫苗株（La Sota）。使用野生型低致病性 lentogenic NDV 株进行 PFU 和 Western blot 实验。
褐藻聚糖对 NDV La Sota 表现出抗病毒活性，获得的 IS50 > 2000。在添加时间研究中，我们观察到病毒在感染的早期阶段受到抑制（感染后 0-60 分钟）。野生型 NDV 株的病毒渗透实验抑制结果支持了这一结果，因为这些实验表明病毒感染降低了 48％，并且 HN 蛋白表达减少。利巴韦林作为抗病毒对照，其抗病毒活性低于褐藻聚糖，且在有效剂量下毒性高。在融合测定中，当褐藻聚糖在融合蛋白切割之前添加时，合胞体的数量明显减少（抑制 70％），这可能表明褐藻聚糖与 F0 蛋白之间存在特异性相互作用。
本研究结果表明，来自 C. okamuranus 的褐藻聚糖是家禽业潜在的低毒性抗病毒化合物，我们的研究结果为硫酸多糖的作用机制提供了更好的理解。
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